Studies On Extraction Of Phospholipid And Purification Of The Phosphatidylcholine Of Pacific Krill

Marine organisms are a good source of phospholipids. and contain a large quantity of polyunsaturated fatty acids (PUFA), especially EPA (eicosapentaenoic acid), DHA (docosahexaenoic acid) and other n-3 series polyunsaturated fatty acids (n-3PUFA, also known as the ω-3 series polyunsaturated fatty acids. ω-3 PUFA).This allows the formation of phospholipids with a special compositions of fatty acid (namely DHA/EPA phospholipids) in the body of marine organisms. In addition to having dual physiological functions of phospholipids and polyunsaturated fatty acids (DHA, EPA), DHA/EPA phospholipids (especially DHA/EPA-PC) has many unique physiological functions because of its combination of unique fatty acid and phospholipid molecules skeleton. For example:the protection of eyesight, improvement of learning ability, prevention of cardiovascular diseases and cancers, increasing of membrane fluidity and so on. Therefore, DHA/EPA phospholipids in the pharmaceutical care has great value of development and utilization.At present, on the market there are two main phospholipids, namely, soybean phospholipid and egg yolk phospholipid. While marine phospholipids rich in DHA, EPA research is relatively few, especially marine PC.This study aimed at developing an industrialization technology of extraction and separation of high quality, low cost, green Pacific krill phospholipids and Pacific krill PC. Begin with study on extraction and purification process of the Pacific krill phospholipids, and then to get the Pacific krill phospholipids as raw materials, study the separation and purification of Pacific krill PC technology and optimization the process Specific contents and results are as follows:Firstly, optimize the conditions of the ethanol extract of the Euphausia pacifica Pacific krill phospholipids by single factor experiment and orthogonal experiment, with the Euphausia pacifical as raw materials and the Pacific krill phospholipids purity and recovery of product as indicators. The best experimental conditions: remove the oil after extract the phospholipids. extraction solvent is 95% ethanol. liquid to solid ratio is 5:1. extraction temperature is 40℃. extraction time is 3h. mix the material by shaking assisted, extraction times is twice. Under the optimum extraction conditions. Pacific krill phospholipids recovery of the product is 90.13%. purity is 28.65%.Secondly, optimize the conditions of oil removal experiment with the Pacific krill phospholipids of the Euphausia pacifical extracted by ethanol as raw material and the Pacific krill phospholipids purity and recovery of product as indicators. The best experimental conditions:Acetone to be de-oiling agent, ratio of liquid to solid is 10:1. ultrasonic time is 25min. ultrasonic power is 250w, ultrasonic temperature is 40℃ and removes the oil 2 times. Under the optimum extraction conditions. Pacific krill phospholipids recovery of the product is 89.26% and the purity increased from 28.65% to 52.93%. Analysis by TLC scanning, the main component and content of Pacific krill phospholipids in the product is 49.68% Pacific krill PC,5.32% PE. while some SM and LPC. The product may further be used to purify Pacific krill phosphatidylcholine by chromatography.Thirdly, optimize the conditions of column chromatography experiments with the product of the above step as raw material and the Pacific krill PC purity and recovery of product as indicators. Because of the small amount of the solvent used and low bands overlap, Al2O3 chosen to be the stationary phase of column chromatography experiments. Optimal conditions for column chromatography:diameter height of column is D25× 300mm, stationary phase is Al2O3 of 100-200 mesh, mobile phase is 95% ethanol, the loading is 0.3g/gAl2O3, flow rate is 3mL/min. Under these conditions, the Pacific krill PC recovery of product is 65.33%, purity is 94.29%.Finally, study the effects of different temperatures, light conditions, air exposure on the oxidation stability of Pacific krill PC. Analysis shows that, low temperature, light avoided and cutting off the air is good for the oxidation stability of Pacific krill PC during storage, in which, the effect of low temperature and cutting off the air is more significant than the dark.